Identification Of MiRNAs Regulates Apoptosis By Targeting PUMA In Lung Cancer

Objective we use Bioinformatics softwares to screen the PUMA relate miRNAs and examed the expression of the selected miRNAs through real time fluorescent PCR in lung cancer, we discussed the relationship between the infection of EB virus and lung cancer, then further Explored the expression of miR-BART5which encode by EB virus in lung cancer.Methods1. We screened Bioinformatics Database to find out the miRNAs that possible regulated the PUMA, then special real time fluorescent PCR test kit has been made.2. the express levels of miRNAs that collect from Lung cancer tissue and lung benign lesion were tested by the particular kid we made, we analyze the Correlation Between individual miRNA pression situation and clinic pathological parameters.3. amplified miRNA which show the significant expression difference, sequenced and analyzed through TA DNA recombinant and sequencing techniques and primly testified the target miRNA sequence.4.we use real time fluorescent PCR to test the EB infection in fress cancer and benign lesions tissue, and exam the expression situation of miR-BART5which encode by EB virus and sequenced and analyzed through TA DNA recombinant and sequencing techniques to testify the Correlation of the target sequence.Results1.96possible regulated PUMA related miRNAs has been screened through the Bioinformatics database, then we customized it into the special kit to test the expression with real time fluorescent PCR. The results show:10types miRNAs were up expression in lung cancer tissue, among them, miRNA-221expression up16times. 2. literature reference in the study of NPC indicate that PUMA was the target gene of miRNA221222, the expression of PUMA was Negative Correlated with it in miRNA221222which were selected for deep analyze. Results with rank sum test show that statistical significance (P<0.05) in lung cancer and Para cancer tissue.3. we tested the infection of EB virus and lung cancer and results show: infection rate of EB virus in21collections of lung cancer was42.85%(9/21) the other11collections infection turn to negative.4.we use real time fluorescent PCR to test the expression situation of miR-BART5which encode by EB virus in Lung cancer tissue and Normal tissue through rank sum test, results indicate that expression of miR-BART5had statistical significance (P<0.05) between Lung cancer and lung benign lesion tissue.5. Successfully connected the amplify products miR-221and miR-BART5to pMD19-T vector, the sequenced results were perfect match the gene bank data.Conclusions1. real time fluorescent PCR test show several types of PUMA related miRNAs up-expressed in tissue lung, which indicate there miRNA have relationship with PUMA in gene express regulation and cancer generation and development.2.Deep research need to made to certified whether PUMA is the target gene of miR-221and miR-BART5, the regulation to PUMA and the relationship with lung cancer.